Brain Tumor Research Center, Beijing Neurosurgical Institute, Beijing Tiantan Hospital affiliated to Capital Medical University, Tiantanxili 6, Chongwen District, Beijing, 100050, People's Republic of China.
Clin Exp Med. 2012 Sep;12(3):135-43. doi: 10.1007/s10238-011-0152-7. Epub 2011 Aug 28.
Gaucher disease is an autosomal recessive lysosomal storage disorder resulting in a deficiency of glucocerebrosidase (GC). Imiglucerase, a recombinant form of GC, has been successfully used in the treatment of Gaucher disease and has been shown to be a good potential candidate for gene therapy. However, its low transduction efficiency and short duration of expression have limited it as a gene therapy strategy. VP22, the herpes simplex virus type I tegument protein, is known to facilitate intercellular protein transport, thus making it a promising tool for improving gene transfer efficiency. To investigate whether the fusion of VP22 to GC could improve its therapeutic efficiency for Gaucher disease, the lentiviral vectors pHIV-GC and pHIV-VP(22)-GC were constructed and confirmed by PCR or RT-PCR. After packaging, the vectors were transduced into human Gaucher II fibroblast cells (GII cells). Flow cytometric analysis revealed that the GC expression rates in lenti-VP(22)-GC-transduced GII cells were higher by comparison than those in lenti-GC-transduced GII cells. A Western blot demonstrated higher levels of GC protein expression in lenti-VP(22)-GC-transduced GII cells. In addition, the long-term expression levels and increased GC activities in lenti-VP(22)-GC-transduced GII cells were also observed. These data implicate that VP22-mediated effects may be useful for enhancing the efficacy of this Gaucher disease treatment.
戈谢病是一种常染色体隐性溶酶体贮积症,导致葡萄糖脑苷脂酶(GC)缺乏。伊米苷酶是 GC 的重组形式,已成功用于戈谢病的治疗,并已被证明是基因治疗的良好候选者。然而,其转导效率低和表达持续时间短限制了它作为基因治疗策略的应用。单纯疱疹病毒 I 衣壳蛋白 VP22 已知可促进细胞间蛋白质转运,因此成为提高基因转移效率的有前途的工具。为了研究 VP22 与 GC 的融合是否可以提高其治疗戈谢病的疗效,构建了 pHIV-GC 和 pHIV-VP(22)-GC 慢病毒载体,并通过 PCR 或 RT-PCR 进行了确认。包装后,将载体转导至人戈谢病 II 型成纤维细胞(GII 细胞)。流式细胞术分析显示,与 lenti-GC 转导的 GII 细胞相比,lenti-VP(22)-GC 转导的 GII 细胞中 GC 的表达率更高。Western blot 显示 lenti-VP(22)-GC 转导的 GII 细胞中 GC 蛋白表达水平更高。此外,还观察到 lenti-VP(22)-GC 转导的 GII 细胞中 GC 活性的长期表达水平增加。这些数据表明 VP22 介导的作用可能有助于增强这种戈谢病治疗的疗效。